公开(公告)号：WO2018064108A1

公开(公告)日：2018-04-05

申请号：PCT/US2017/053611

申请日：2017-09-27

Applicant: INVISTA NORTH AMERICA S.Á.R.L.

Inventor： BAWDON, Daniel ,  CARTMAN, Stephen, Thomas ,  KENNEDY, Jonathan

IPC: C12N15/74 ,  C12N9/22

Abstract: Nonnaturally occurring host cells altered to increase their ability to transfer genetic molecules into the host cells as compared to an unaltered host cell are provided. Also provided are methods for identifying endogenous loci of a host cell which inhibit transformation efficiency and/or electroporation of genetic molecules into the cell as well as methods for producing nonnaturally occurring host cells with enhanced transformation efficiency and/or the modified ability to allow for genomic integration of an exogenous DNA sequence via electroporation. Methods for producing biochemicals and products produced with the nonnaturally occurring host cells are also provided. In particular, a host Cupriavidus necator cell is provided in which a chromosomal gene encoding a restriction endonuclease has been disrupted, leading to an increase in transformation efficiency.

Abstract translation: 与未改变的宿主细胞相比，非天然存在的宿主细胞被改变以提高其将遗传分子转移到宿主细胞中的能力。 还提供了用于鉴定抑制转化效率和/或遗传分子电穿孔进入细胞的宿主细胞的内源基因座的方法，以及用于产生具有增强的转化效率和/或修饰的基因组学能力的非天然存在的宿主细胞的方法 通过电穿孔整合外源DNA序列。 还提供了生产生物化学品和用非天然存在的宿主细胞产生的产物的方法。 具体而言，提供了宿主Cupriavidus necator细胞，其中编码限制性内切核酸酶的染色体基因已被破坏，导致转化效率提高。